The Universal Primers and the Shotgun DNA Sequencing Method
For studies in molecular biology, DNA purification has been essential, in particular for DNA sequencing, probing, and mutagenesis. The amplification of DNA in Escherichia coli by cloning vehicles derived from M13mp or pUC made expensive physical separation techniques such as ultracentrifugation unnecessary. Although today the polymerase chain reaction (PCR) is a valuable alternative for the amplification of small DNA pieces (1 ), it cannot substitute for the construction of libraries of DNA fragments. Therefore, E. coli has served not only as a vehicle to amplify DNA, but also to separate many DNA molecules of similar length and the two DNA strands simultaneously. For this purpose, a bacteriophage such as M13 can be used. The various viral cis - and trans -acting functions are critical not only for strand separation, but also to separate the single-stranded DNA from the E. coli cell by an active transport mechanism through the intact cell wall.
- Cytokine Gene-Modified Cell-Based Cancer Vaccines
- Epitope Mapping by Surface Plasmon Resonance
- Single-Marker Family-Based Association Analysis Conditional on Parental Information
- Using Nuclear Run-On Transcription Assays in RNAi Studies
- DNA Extraction of Ancient Animal Hard Tissue Samples via Adsorption to Silica Particles
- Generation of Random Fragments by Sonication
- Assessing Sperm DNA Fragmentation with the Sperm Chromatin Dispersion Test
- Cloning, Expression, and Functional Analysis of Genomic miRNA Using Retroviral System in Cancer Cells
- Laboratory Methods for the Analysis of Primate Mobile Elements
- Association Mapping Using Pooled DNA